Hematoxyllin-Eosin ( HE ) Staining
Freda Halim
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Abstract
This is a protocol to stain tissue with Hematoxyllin-Eosin ( HE ) Staining. We use human breast cancer tissue for running this protocol.
Steps
Deparaffinization
Incubate slides in Xylol
Incubate slides in Xylol
Rehydration
Incubate slides in 100% Alcohol
Incubate slides in 96% Alcohol
Incubate slides in 80% Alcohol
Incubate Slides in 70% Alcohol
Rinse slides with Aquadest for 5 minutes
Nuclear Staining
Incubate Slides in Hematoxyllin Reagent ( Hematoxyllin JT Baker )
Rinse slides with running tap water
Defferentiation
Incubate slides in bluing reagent @ 1 minutes, 5 times
Rinse slides with running tap water
Cytoplasm staining
Incubate slides in Eosin reagent 1 minute
Dehydration
Incubate Slides in 70% Alcohol
Incubate Slides in 80% Alcohol
Incubate Slides in 96% Alcohol
Incubate Slides in 100% Alcohol
Clearing
Incubate Slides in Xylol
Incubate slides in Xylol
Incubate Slides in Xylol
Mounting and Finishing
Wipe excess fluid,cover the slides with cover glass